Fig. 3

Verification and characterization of OE::FfH2A.Z and TetOff::FfH2A.Z strains in Fusarium fujikuroi (FfWT). (A) Expressional analysis of the FfH2A.Z mutant strains OE::FfH2A.Z and TetOff::FfH2A.Z via RT-qPCR. For the FfH2A.Z depletion strain, FfH2A.Z expression was measured upon cultivation on CM supplemented with (50 µg/mL) and without (0 µg/mL) the inducing reagent doxycycline hyclate (DOX). The FfH2A.Z overexpression strain was cultivated on CM at 30 °C for 3 days in the dark. Gene expression of FfWT was arbitrarily set to 1. (B) Western blot analysis of FfWT, FfH2A.Z knock-down and overexpression mutant strains. Proteins were probed with an anti-H2A.Z-specific and an anti-H3 C-terminus-specific antibody. For quantification, a densitometric analysis was performed where FfWT was arbitrarily set to 1. (C) Assessment of radial hyphal growth of the inducible TetOff::FfH2A.Z strains and FfWT on CM using different concentrations of the inducing agent DOX. The medium was inoculated either with a mycelial plug (left panel) or 1,000 conidia each (right panel) and incubated for 4 days post inoculation at 30 °C in the dark. Experiments were performed in biological triplicates